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Limit of Detection (LOD)
Lowest amount of allergen to be distinguished from a true blank
LOD mostly determined in buffer
May be completely different in food samples
Recovery
Ideally 80 – 120%
Not possible for all matrices
(pH, salts, polyphenols,…)
Spiked vs. incurred samples
Factors influencing ELISA comparability
Matrix Choices: Sources of Interference
•
High protein concentration
•
High lipid concentration
•
High polysaccharide concentration
•
Extremes of pH
•
May push past pH-buffering capacity of kit reagents
•
Reactive biomolecules
•
Polyphenols
•
Tannins
•
Anthocyanins
Should choose matrices which test ability of the
assay to buffer sources of interference